Enterovirus sequencing as a new approach to the laboratory diagnosis for clinical and epidemiological purposes
Abstract
Objective: Introducing sequencing of enteroviruses as progressive methods for the classification of viruses isolated according to the new nomenclature and detailed assessment of the nature of the isolate. Materials and Methods: It was analyzed 75 samples drawn from 64 patients from two departments - Regional Hospital in Liberec and Plzeň Teaching Hospital.
Age of the patients ranged from 4 to 54 years, the median age was 15 years for men and 16 years for women. The reason for hospitalization were the predominant number of patients, intense headache, fever, vomiting, fatigue, meningeal symptoms, two patients were bowel symptoms and a skin.
Investigated samples were swabs from the rectum, throat, cerebrospinal beverages of a stool. Molecular detection and typing was performed using RT-PCR method.
For typing was chosen stretch of non-coding region 5'RNA. Samples were amplified by one-step PCR with outside primers and the same primers extended on M13 sequences (Generi-Biotech).
Editing, assembly and qualitative analysis of the sequences was performed using the software Lasergene (DIASTAR). The sequences obtained were evaluated by the central database GenBank sequences using the BLAST algorithm.
The results: In the isolates we identified 61 ECHO virus 30 3 coxsackieviry B1 1 coxsackievirus B3 1 coxsackievirus A9 1 enterovirus 86, enterovirus 71 1, 2 ECHO virus 13 / coxsackievirus B5 1 ECHO virus 7/30 / coxsackievirus B4, 1 coxsackievirus B4 / enterovirus B, enterovirus 87 1 / ECHO virus 30 / enterovirus B 1 and ECHO virus 3. All isolated viruses, including enterovirus 71 belonging to group A belong to the group B.
CONCLUSION: Sequencing of fully replacing closer determination of enteroviruses Virus neutralization and allow a detailed assessment of the isolates. The predominance of ECHO 30 as the causal agent causing aseptic meningitis is consistent with the literature.